Osmani S A, May G S, Morris N R
J Cell Biol. 1987 Jun;104(6):1495-504. doi: 10.1083/jcb.104.6.1495.
The temperature-sensitive cell cycle mutation nimA5 causes nuclei of Aspergillus nidulans to be blocked in late G2 at restrictive temperature. Under these conditions the spindle pole body divides but does not separate and the mitotic index drops to zero. If nimA5 is blocked for more than one doubling time and then shifted from restrictive to permissive temperature, nuclei immediately enter mitosis, the mitotic spindle forms, and the chromosomes condense (Oakley, B. R., and N. R. Morris, 1983, J. Cell Biol., 96:1155-8). We have cloned the wild-type nimA gene by DNA-mediated complementation of the nimA5 mutant phenotype and have characterized nimA mRNA expression by Northern blot analysis. The transcript is 3.6 kb in length and is under tight nuclear cycle regulation. In synchronously dividing cells, the levels of nimA mRNA become elevated as cells enter mitosis and drop sharply as cells progress through mitosis. Cells blocked in S-phase with hydroxyurea have very low levels of nimA mRNA. Cells blocked in mitosis, either by the antimitotic agent benomyl or by the cell cycle mutation bimE7, maintain elevated levels of the nimA transcript. These data demonstrate not only that nimA is required for entry into mitosis, but because the transcript is normally expressed cyclically and is under tight cell cycle control, they suggest that nimA may play a regulatory role in the initiation of mitosis.
温度敏感型细胞周期突变体nimA5会使构巢曲霉的细胞核在限制温度下阻滞于G2晚期。在这些条件下,纺锤极体分裂但不分离,有丝分裂指数降至零。如果nimA5被阻滞超过一个倍增时间,然后从限制温度转移至允许温度,细胞核会立即进入有丝分裂,有丝分裂纺锤体形成,染色体凝聚(奥克利,B.R.,和N.R.莫里斯,1983,《细胞生物学杂志》,96:1155 - 1158)。我们通过对nimA5突变体表型的DNA介导互补克隆了野生型nimA基因,并通过Northern印迹分析对nimA mRNA表达进行了表征。该转录本长度为3.6 kb,处于严格的核周期调控之下。在同步分裂的细胞中,随着细胞进入有丝分裂,nimA mRNA水平升高,而随着细胞进行有丝分裂,其水平急剧下降。用羟基脲阻滞在S期的细胞中nimA mRNA水平非常低。无论是被抗有丝分裂剂苯菌灵还是细胞周期突变体bimE7阻滞在有丝分裂期的细胞,都维持nimA转录本的高水平。这些数据不仅证明进入有丝分裂需要nimA,而且因为该转录本通常呈周期性表达且处于严格的细胞周期控制之下,所以表明nimA可能在有丝分裂起始中发挥调节作用。